Phospho-IRS1 (Ser312) Polyclonal Antibody

Phospho-IRS1 (Ser312) Antibody (44-814G)

Peptide Competition and Phosphatase Stripping. Extracts of CHO-T cells transiently transfected with wild-type human IRS-1 and treated with 100 ng/mL TPA for 30 minutes were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C and either left untreated (2-6) or treated with lambda phosphatase (1), then incubated with the IRS-1 [pS312] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1-3), the non-phosphopeptide corresponding to the phosphopeptide immunogen (4), a generic phosphoserine-containing peptide (5), or the phosphopeptide immunogen (6). After washing, the membrane was incubated with goat F(ab&quote;)2 anti-rabbit IgG alkaline phosphatase (cat. ## ALI4405) and signals were detected using the Tropix WesternStar™ method. The data show that only the phosphopeptide corresponding to IRS-1 [pS312] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.

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Product Details

Bioinformatics

Documents

The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human IRS-1 that contains serine 312. The sequence is conserved in mouse, rat and pig.

Conjugate

Unconjugated

Form

Liquid

Purification

Antigen affinity chromatography

Storage buffer

Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol

Contains

0.05% sodium azide

Storage Conditions

-20°C

Tested Applications

Dilution *

Western Blot (WB)

Assay Dependent

* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the
product for use in their own experiment using appropriate negative and positive controls.

Background/Target Information

Insulin receptor substrates (IRS) are responsible for several insulin related activities, such as glucose homeostasis, cell growth, cell transformation, apoptosis and insulin signal transduction. Serine/threonine phosphorylation of IRS-1 has been demonstrated to be a negative regulator of insulin signaling and is responsible for its degradation, although IRS-1 degradation pathways are not well understood. IRS-1 has also been shown to be constitutively activated in cancers such as breast cancer, Wilm&quote;s tumors, and adrenal cortical carcinomas, thus making IRS-1 phosphorylation and subsequent degradation an attractive therapeutic option. To date there have been four subtypes identified: IRS-1,2,3, and 4, with IRS-1 being widely expressed.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.